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Merck

CGP1

Sigma-Aldrich

Glutathione Peroxidase Cellular Activity Assay Kit

Sufficient for 100 colorimetric tests

Sinónimos:

Cellular glutathione peroxidase, c-GPx

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About This Item

Código UNSPSC:
12161503
NACRES:
NA.32
En este momento no podemos mostrarle ni los precios ni la disponibilidad

Nivel de calidad

uso

sufficient for 100 tests

caducidad

Kit is stable for 24 months when unopened.

método de detección

colorimetric

Condiciones de envío

wet ice

temp. de almacenamiento

−20°C

Descripción general

The Glutathione Peroxidase Cellular Activity Assay kit is used to measure glutathione peroxidase in tissue extracts. The assay is based on the oxidation of glutathione (GSH) to oxidized glutathione (GSSG). This is catalyzed by GPx coupled to the recycling of GSSG back to GSH utilizing glutathione reductase and NADPH. The decrease in NADPH absorbance measured at 340 nm during the oxidation of NADPH to NADP is indicative of glutathione peroxidase activity since the enzyme is the rate-limiting factor of the coupled reactions.

Idoneidad

Suitable for the measureent of glutathione peroxidase in tissue extracts

Principio

The Glutathione Peroxidase Cellular Activity Assay kit is used to measure glutathione peroxidase in tissue extracts. The assay is based on the oxidation of glutathione (GSH) to oxidized glutathione (GSSG). This is catalyzed by GPx coupled to the recycling of GSSG back to GSH utilizing glutathione reductase and NADPH. The decrease in NADPH absorbance measured at 340 nm during the oxidation of NADPH to NADP is indicative of glutathione peroxidase activity since the enzyme is the rate-limiting factor of the coupled reactions.

Otras notas

This kit was tested with rabbit reticulocyte lysate and with glutathione peroxidase enzyme.

Solo componentes del kit

Referencia del producto
Descripción

  • tert-Butyl hydroperoxide 1 mL

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Palabra de señalización

Danger

Clasificaciones de peligro

Acute Tox. 2 Inhalation - Acute Tox. 3 Dermal - Acute Tox. 4 Oral - Aquatic Chronic 2 - Eye Dam. 1 - Flam. Liq. 3 - Muta. 2 - Org. Perox. F - Skin Corr. 1C - Skin Sens. 1 - STOT SE 3

Órganos de actuación

Respiratory system

Código de clase de almacenamiento

5.2 - Organic peroxides and self-reacting hazardous materials


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V R Arruda et al.
Neoplasma, 43(2), 99-102 (1996-01-01)
Red cell antioxidant enzymes have been recently studied in malignant lymphomas and the results are controversial. Hairy cell leukemia is a rare chronic lymphoproliferative disorder originating probably in a pluripotent stem cell. In the present study, glutathione peroxidase (Gpx), reduced
Ian M Scott et al.
Pesticide biochemistry and physiology, 141, 9-17 (2017-09-16)
Plant elicitors can be biological or chemical-derived stimulators of jasmonic acid (JA) or salicylic acid (SA) pathways shown to prime the defenses in many crops. Examples of chemical elicitors of the JA and SA pathways include methyl-jasmonate and 1,2,3-benzothiadiazole-7-carbothioate (BTH
International Committee for Standardization in Haematology: recommended methods for red-cell enzyme analysis.
E Beutler et al.
British journal of haematology, 35(2), 331-340 (1977-02-01)
Diversity of glutathione peroxidases.
F Ursini et al.
Methods in enzymology, 252, 38-53 (1995-01-01)
Roxana Oana Cojocariu et al.
Brain sciences, 10(11) (2020-11-21)
Background and Objectives: Irritable bowel syndrome (IBS) is a well-known functional gastrointestinal (GI) disorder exhibiting a wide range of symptoms due to individual variability and multifactorial etiology. Stress exposure is a major risk factor for the development of IBS. Here

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Cellular oxidative stress is countered by enzymatic scavengers and antioxidant modulators against reactive oxygen species damage.

Cellular oxidative stress is countered by enzymatic scavengers and antioxidant modulators against reactive oxygen species damage.

Cellular oxidative stress is countered by enzymatic scavengers and antioxidant modulators against reactive oxygen species damage.

Cellular oxidative stress is countered by enzymatic scavengers and antioxidant modulators against reactive oxygen species damage.

Questions

1–8 of 8 Questions  
  1. "Does A340/MIN refer to the absorbance value per minute, or is it the rate of decrease per minute?"

    1 answer
    1. The CGP1 assay measures the decrease in NADPH absorbance at 340 nm during the oxidation of NADPH to NADP+. This is a kinetic assay and the A340/MIN means absorbance per minute. Please see step 4 on page 4 of the technical bulletin for the kinetic program instructions. The number of readings being 6 for an interval time of 10 seconds which means data is for a minute. NADP+ does not have any absorbance at 340 nm and NADPH does. Thus, the absorbance will decrease in a sample with glutathione peroxide as all substrates, cofactors, enzymes are present for this conversion to occur.

      Helpful?

  2. Can Product CGP1, Glutathione Peroxidase Cellular Activity Assay Kit, be used with Serum samples?

    1 answer
    1. The Glutathione Peroxidase Cellular Activity Assay Kit (Product number: CGP1) is designed to measure Glutathione Peroxidase activity in cell lysates and tissue homogenates. This kit has not been validated for use with serum samples.

      Please see product MAK437, Glutathione Peroxidase Assay Kit, which can be used for serum samples:
      https://www.sigmaaldrich.com/product/sigma/mak437

      Helpful?

  3. Can Product CGP1, Glutathione Peroxidase Cellular Activity Assay Kit, be run in a 96 well plate?

    1 answer
    1. Yes, but you would need to have a plate reader that has a kinetic function as this is a kinetic assay.

      Helpful?

  4. What dilution should I use for my sample when using Product CGP1, Glutathione Peroxidase Cellular Activity Assay Kit?

    1 answer
    1. In general, when human erythrocytes were assayed for the enzyme, the sample was diluted 10-fold (15 mg peroxidase per mL) and a 20 μl sample gave a decrease of 0.032 OD340 per minute. Too much enzyme will underestimate the apparent activity of the sample. When rabbit reticulocytes were assayed at a 10-fold dilution (10 mg peroxidase per mL), 20 μl gave a decrease of 0.065 OD340 per minute.

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  5. What is the Department of Transportation shipping information for this product?

    1 answer
    1. Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.

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  6. What type of readings do I get with Product CGP1, Glutathione Peroxidase Cellular Activity Assay Kit?

    1 answer
    1. There is a decrease in absorbance during the assay. The OD values decrease with the increase in reaction time. The slope of the reduction in OD (delta OD per minute) is the value used to determine the enzyme units.

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  7. How should I prepare my sample for analysis with Product CGP1, Glutathione Peroxidase Cellular Activity Assay Kit?

    1 answer
    1. The cells must be lysed before assaying with this kit.  The best way to perform lysis is to incubate the cells with hypotonic buffer (at least 2 volumes and better with 3-4 volumes, depending on the expected level of peroxidase in these cells). Let them swell and homogenize with a Dounce homogenizer. Centrifuge the homogenate at 10,000 × g and use the supernatant for the assay.  If a detergent is used for lysis, only use a peroxide-free detergent and follow the instructions in the Bulletin: "Nonionic detergents such as TWEEN and TRITON X-100 that contain high levels of endogenous peroxides will raise the apparent activity. If these detergents are vital to the extraction of the proteins of interest, a low peroxide detergent should be used such as Product Codes X-100-PC (TRITON X-100), P 6585 (TWEEN 20), or P 8192 (TWEEN 80)".

      Helpful?

  8. Can Product CGP1, Glutathione Peroxidase Cellular Activity Assay Kit, be used with tissue samples?

    1 answer
    1. We have not tested the CGP1 assay with tissue samples. However, we suggest that the customer perform a small-scale pilot experiment using our CelLytic-MT lysis buffer (Product No. C3228). The buffer enables efficient extraction of tissue proteins. It does not contain any of the interfering agents that are listed on the kit bulletin.

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